Morphological Variation in Eight Taxa of Anthyllis Vulneraria S. Lato (Fabaceae)

Ann. Bot. Fennici 42: 293–304 ISSN 0003-3847 Helsinki 30 August 2005 © Finnish Zoological and Botanical Publishing Board 2005

Morphological variation in eight taxa of Anthyllis vulneraria s. lato (Fabaceae)

Egle Puidet, Jaan Liira, Jaanus Paal, Meelis Pärtel & Silvia Pihu

Institute of Botany and Ecology, University of Tartu, 40 Lai Str., 51005 Tartu, Estonia

Received 5 Oct. 2004, revised version received 10 Dec. 2004, accepted 17 Dec. 2004

Puidet, E., Liira, J., Paal, J., Pärtel, M. & Pihu, S. 2005: Morphological variation in eight taxa of Anthyllis vulneraria s. lato (Fabaceae). — Ann. Bot. Fennici 42: 293–304.

Depending on the literature source, the number of existing Anthyllis species differs almost three-fold. In addition to the well-defined species, there are many cryptic ones. Statistical analysis (general linear models, discriminant analysis) of the morphological variation of eight Anthyllis taxa (for simplification classified as species) resulted in three groups of species: Vulneraria (A. vulneraria, A. maritima, A. arenaria and A. ¥ baltica), Macrocephala (A. macrocephala, A. ¥ colorata, and A. ¥ polyphylloides), and Coccinea (A. coccinea). Distinguishing features of these groups were calyx colour, corolla colour, hairiness of stems and petioles, and plant height.

Key words: Anthyllis vulneraria, morphology, suboptimal classiﬁcation, taxonomy, variation

Introduction tion (Yakovlev et al. 1996). The species number has been given as 25 (Cullen 1986) up to 60 The genus Anthyllis (Fabaceae) is one of eight (Minjaev & Akulova 1987). Although some spe- genera in the tribe Loteae and is morphologically cies in the genus are well defined and universally and molecularly closely related to the genus accepted, there are many cryptic forms that have Hymenocarpus (Polhill 1994). This relationship been subject to different interpretations. The spe- has been confirmed in later studies (Allan & cies are morphologically quite similar. Although Porter 2000, Allan et al. 2003), which substanti- the genus contains some shrubs and subshrubs, ate the sister group relationship between Anthyl- all European species are herbaceous (Cullen lis and Hymenocarpus xerxinnatus. 1968). Cullen (1968) divided A. vulneraria s. Anthyllis vulneraria s. lato occurs from lato into three groups: subsp. vulneraria, subsp. the Volga River to England and from northern maritima and subsp. polyphylla. Europe to the Mediterranean (Hultén & Fries There are two main schools of thought with 1986a). It has also been introduced into North respect to the taxonomy of the genus Anthyl- America and New Zealand (Hultén & Fries lis. The first, popular in the area of the former 1986b). The exact number of Anthyllis species Soviet Union, distinguishes numerous species is controversial and depends on interpretation in Anthyllis vulneraria s. lato (Juzepczuk 1945, of their morphological-geographical boundaries Minjaev & Akulova 1987). The other line of with respect to active speciation and hybridisa- thought, prevalent in most of Europe, recognises 294 Puidet et al. • ANN. BOT. FENNICI Vol. 42

18 European species (Cullen 1968, Garcke 1972, detectable characteristic is hair disposition on the Hegi 1975, Ulvinen & Lampinen 1998) with stem and petiole. Anthyllis macrocephala, A. ¥ many species of the first school being recognised polyphylla and A. ¥ colorata have patent hairs on as subspecies or varieties. the stems and petioles, whereas the other species Anthyllis vulneraria s. lato is a particularly have appressed hairs (Cullen 1968, Garcke 1972, polymorphic taxon (Krall 1983). Its intraspecific Hegi 1975, Krall 1983, Eglite & Krall 1996, Krall classification is complicated; subspecies and 1999). Anthyllis maritima can be distinguished forms are closely similar and often have a hybrid from the other species by concolorous calyces, origin. Anthyllis vulneraria consists of local forms by sericeous calyx pubescence and some inflo- in limited areas, which are almost morphologi- rescences with few flowers (sometimes not fully cally indistinct (Jalas 1950, Talts 1959). On old developed) (Krall 1983, Eglite & Krall 1996, Roze arable lands in central and western Europe, hybrid 2004). Inflorescences of this species also feature complexes whose taxonomic status is difficult to long peduncles. Anthyllis arenaria has well-devel- determine have been found (Talts 1959). Estonian oped inflorescences that are sessile (Eglite & Krall habitats are similar to those in central and southern 1996, Krall 1999). Branches of this species form Sweden, where several varieties of Anthyllis have an acute angle with the stem (Roze 2004). Of been described and where populations of Anthyllis the species with bicoloured calyces, A. coccinea occur typically as hybrid complexes (Jalas 1950). is most readily distinguished by its red corolla This article investigates eight taxa (A. are- (Cullen 1968, Krall 1983, Eglite & Krall 1996, naria (Rupr.) Juz., A. coccinea (L.) Beck, A. Krall 1999). Anthyllis ¥ baltica has also some macrocephala Wend., A. maritima Schweigg. undeveloped inflorescences in axils, like A. mar- and A. vulneraria L., s. stricto, A. ¥ colorata itima (Eglite & Krall 1996, Roze 2004). Anthyllis Juz., A. ¥ baltica Juz., A. ¥ polyphylloides Juz.), vulneraria s. stricto has unbranched stems and which are recognised as species in this study, mainly apical inflorescences (Juzepczuk 1945, as they are also in the Baltic States (Talts 1959, Eglite & Krall 1996, Krall 1999). According to the Krall 1983, Eglite & Krall 1996, Krall 1999, keys and floras these eight taxa can be readily dis- Kukk 1999). These species and their equivalents tinguished, yet individual plants of genus Anthyl- in other classifications are listed in Table 1. lis in natural stands are difficult to identify. Whereas these eight taxa are considered sub- There are also many infrequently used char- species or varieties of Anthyllis vulneraria s. acteristics that can be found in other studies lato in Europe, their global distribution remains (Cullen 1968, Lukaszewska et al. 1983a, 1983b, indeterminable. The most widespread taxa in the Krall 1983, Tihomirov & Sokoloff 1996). Baltic region, A. vulneraria s. stricto, A. macro- Becker (1912) distinguished two growth cephala, A. maritima and A. arenaria, are distrib- forms in Anthyllis vulneraria s. lato: Vulgaris uted throughout the area, whereas A. coccinea and Vulneraria. They usually grow in similar occurs only in western Estonia and Latvia, and in conditions, but Vulgaris prefers moister habitats southern Sweden (Tabaka 1982, Minjaev & Aku- than Vulneraria. He also claimed that corollas lova 1987, Tabaka et al. 1988). Anthyllis vulner- are usually yellow in moister conditions, but aria and A. macrocephala also occur in southern principally red in dryer habitats. Becker’s results Finland (Ulvinen & Lampinen 1998). Anthyllis contradict all traditionally used characteristics to ¥ polyphylloides has the same distribution as its distinguish Anthyllis species. probable parent species. Anthyllis ¥ baltica is Lukaszewska et al. (1983a, 1983b, 1983c) ana- endemic to the Baltic region and A. ¥ colorata is lysed the morphological variability and Kalinowski endemic to Estonia (Minjaev & Akulova 1987). (1983a, 1983b) the isoenzymatic variability of A. A few characteristics in different keys and vulneraria s. lato populations from coastal areas floras distinguish these taxa (Table 2). Bicol- of the Baltic Sea in Poland. Different methods of oured rufous calyx teeth demarcate A. vulner- multivariate statistical analysis all confirm the dif- aria s. stricto, A. coccinea, A. ¥ baltica and A. ferences between populations, both in vegetative ¥ colorata from the other four species, which and sexual characteristics. For most traits a cor- have concolorous, green calyces. Another readily relation was found between the differentiation of ANN. BOT. FENNICI Vol. 42 • Morphological variation in eight taxa of Anthyllis vulneraria 295 A. (DC.) Sag. Lampinen 1998 vulneraria, polyphylla . linnaei – ssp. Finland Ulvinen & ssp – – Nyman – – – (Schweiggrer) Koch. Koch. vulneraria (Kerner) Wohlf., vulgaris ssp. coccinea L. maritima polyphylla (Kit.) Ser.

Europe Hegi 1975 var. var. – var. – – – A. et G. (Kit.) Arcang. var. L.,* (Schweigg.) vulneraria maritima polyphylla . vulgaris (Koch) A. et G. var.

Europe Gracke 1972 – ssp. ssp – ssp. – Juz. according to different taxonomic arrangements (– = taxon was not cited). *All infraspeciﬁc taxa are members of members are taxa infraspeciﬁc *All cited). not was taxon = (– arrangements taxonomic different to according Anthyllis (Rupr.) Juz. (Meinsh.) Juz. –

Former Soviet Union Juzepczuk 1945 A. coccinea (L.) Beck A. arenaria – A. maritima Schweigg. A. polyphylla Kit. ssp. A. colorata A. polyphylloides

– Miniaev Juz. Wend. . ex

L., s. stricto A. Linnaei Sag. Juz. (L.) Beck

Schweigg. Juz. (Rupr.) Juz. s. lato

colorata polyphylloides baltica Kloczkova

¥ ¥ ¥

Classiﬁcation of species of the genus the of species of Classiﬁcation 1. Table vulneraria Baltic countries Eglite & Krall 1996 A. vulneraria A. coccinea A. arenaria A. maritima A. macrocephala A. A. A.

et 296 Puidet et al. • ANN. BOT. FENNICI Vol. 42 polyphylloides ¥ 3–5 2–4 colorata

¥ erect 2–3 as calyx sharp as calyx 2–3 sharp yellow yellow baltica ¥ 2–3 simple leaves 3–5 3–5 as calyx calyx 1–3 3–4 simple leaves 2–5 2–4 blunt-ended 1–4 yellow yellow yellow yellow species (Krall 1983, 1999, Eglite & Krall 1996). decumbent 1–2 simple leaves 1–2 red Anthyllis vulneraria, s. stricto coccinea 15–30 cm arenaria ascending, 4–15 cm erect ascending, maritima 20–50 cm erect 20–40 cm macrcocephala 16–40 cm ascending 20–40 cm erect as long as calyx 20–50 cm ascending sharp or as long blunt-ended 15–30 cm as calyx ascending, blunt-ended as long as calyx sharp or erect bi-coloured with red teeth longer than calyx bi-coloured sharp with red teeth yellow green as long concolorous, concolorous, sharp concolorous, longer than green, hairy bi-coloured with green bi-coloured as long sharp concolorous, red teeth, hairy as long with red teeth green Main characteristics of the Estonian Table 2. Characteristics Plant height Stem shape Hair disposition on stem and petiole appressed Number of stem leaves appressed Shape of the rosette leaves 2–3 pinnate appressed Length of bracts appressed might be patent Apexes of bracts pinnate Number of inﬂorescences appressed might be 2–3 Colour of the calyx patent pinnate Colour of corolla patent might be pinnate pinnate ANN. BOT. FENNICI Vol. 42 • Morphological variation in eight taxa of Anthyllis vulneraria 297 populations and their geographic locations. sity of Tartu (TU), the Herbarium of the Institute Chromosome numbers have been determined of Zoology and Botany of the Estonian Agricul- only for A. maritima, A. vulneraria and A. mac- tural University (TAA) and the Herbarium of the rocephala, all of which have 2n = 12 chromo- Estonian Natural Museum (TAL). In addition, somes (Tshehov 1932, Jalas 1950, Bakšay 1959, the analysis included 300 specimens material Bolkohovskikh et al. 1969, Agapova et al. 1990), collected from different localities in Estonia in suggesting they are diploid. July 2001 and June 2002. Eight Anthyllis species As much as the determination of Anthyllis occurring in northern Europe and widespread in species and their hybrids remains problematic, the Baltic region were analysed: A. vulneraria the aims of this study were: (i) to determine (86 individuals), A. coccinea (45), A. arenaria the circumscription of the Anthyllis species, (ii) (81), A. maritima (95), A. macrocephala (25), to analyse their variation patterns and, (iii) to A. ¥ baltica (105), A. ¥ colorata (39) and A. ¥ determine which morphological characteristics polyphylloides (22). distinguish these taxa reliably. Twenty-three diagnostic characteristics were selected for analysis (Table 3). These characteristics are those commonly used to identify Material and methods species and those measurable in the herbarium material. The analysis comprised herbarium material (198 The length of the longest stem was designated individuals) from the Herbarium of the Univer- as the height of the plant. The number of stem

Table 3. Characteristics used in analysis of Anthyllis species (degree of precision for metric interval characteristics 1 mm).

Abbreviation Variable Type Scale

SR habit nominal 1 = erect, 2 = ascending HLT apexes of bracts nominal 1 = sharp, 2 = blunt-headed HP hair of calyx nominal 1 = oppressed, 2 = silky, patent hairs SRL shape of rosette leaves nominal 1 = simple leaves, 2 = pinnate HSP hair of petiole nominal 1 = no hairs, 2 = oppressed, 3 = patent HST hair of stem nominal 1 = no hairs, 2 = oppressed, 3 = patent CIF colour of corolla nominal 1 = yellow, 2 = orange, 3 = red DIF branching of inﬂorescence nominal 1 = simple, 2 = ramiform without axes, and without axes 3 = ramiform with axes, 4 = ramiform with HLB hair on the upper side ordinal 1 = no hairs, < 2 = few hairs (1–3 on of leaf blade 2.5 mm2), > 3 = many hairs (3 on 2.5 mm2) CP colour of calyx nominal 1 = concolorous, 2 = some red colour teeth on teeth, 3 = clearly bi-coloured, with red NS number of stems interval counted NIF number of inﬂorescence interval counted NSL number of stem leaves interval counted NRL number of rosette leaves interval counted L height of the plant interval metric (cm) LL length of the leaf interval metric (cm) WL width of the leaf interval metric (cm) LL/WL ratio of the leaf length and width LP length of calyx interval metric (cm) WP width of the calyx interval metric (cm) LP/WP ratio of the calyx length and width LF length of the corolla interval metric (cm) LHL length of bract interval metric (cm) WHL width of bract interval metric (cm) LHL/WHL ratio of bract length and width LS length of the petiole interval metric (cm) 298 Puidet et al. • ANN. BOT. FENNICI Vol. 42

leaves on the longest stem was also counted. The hairiness of the leaf blade was estimated from the biggest rosette leaf (if absent, from the biggest stem leaf). Hairs in the middle of the right

colorata part of the leaf blade were counted in an area of ¥ 2

A. 2.5 mm with a binocular microscope. The length and width of the same leaf were measured. The length and width of the corolla, calyx and bracts were measured on the same inﬂorescence. The

degree of measurement precision was 1 mm. In addition, ratios of length and width were calcu- baltica ¥ lated for the leaves, bracts and calyces. LHL, WHL, LNRL LNSL, LNRL A. L, LL, WL, LP, WP, L, LL, LP, WP, LNIF,

Data analysis

General linear model (GLM, StatSoft Inc. 2001)

A. macrocephala A. No differences WL, LNSL was used to analyse metric interval and counted interval characteristics and their differences among species. Counted interval characteristics were log-transformed. As the second step in GLM analysis, Tukey’s HSD test was used to estimate which species differ statistically signiﬁ- WHL, LNIF A. maritima cantly in respect to the characteristics indicated by GLM results. Nominal and ordinal characteristics were

analysed with a nonparametric test (Kruskal- Wallis ANOVA). Discriminant analysis (StatSoft Inc. 2001) LNIF A. arenaria was used to determine the subset of character-

< 0.05) according to GLM analysis. Characteristic abbreviations as in Table 3. istics that reliably distinguish the species. Data p were analysed by standard discriminant analysis

methods including non-transformed data. A classification matrix was calculated to evaluate the results. To achieve a suboptimal classification, itera- A. coccinea L, LL, WL, LP, WP, LNS, LNIF, LNSL L, LL, WL, LP, LF, LHL, LS, LNS, LNIF, LNSL No differences L, LL, WL, LP, WP, LF, LHL, WHL, LS, LNIF, LNSL No differences No differences LNIF, LNSL LNIF, LNSL L, LL, WL, WP, LP, L, LL, WL, LNIF L, LL, WL, LP, WP, tive L, LNIF discriminant analysis (StatSoft Inc. 2001) was performed, in which the initial classification was iteratively corrected according to the posterior probabilities until all the specimens were 100% correctly reclassified according to the classification matrix. Next, canonical discriminant analysis was performed and classification similar- LS, LNS, LNIF, LNSL LNIF, LNSL A. vulneraria L, LL, WL, LF, LS, LNSL L, LL, WL, LP, WP, LHL, WHL, LS, LNS, LNIF L, LL, WL, LP, LHL, LS, LNIF, LF, LHL, WHL, LNSL LHL, WHL, LS, LNIF LNIF L, LS, LNS, LNIF, LNSL, LNRL L, LL, WL, LNS, L, LL, WL, LS, LNS, LL, WL, LP, WP, L, LL, WL, LS, LNS, LL, LNS, LNIF No differences LNSL L, LL, LP, LNS WP, LHL, WHL, ity LHL, WHL, LS, LNS, between groups of preliminary species classification and reclassified species were calculated in a frequency matrix. Canonical discriminant analysis (StatSoft Inc. 2001) was used for ordination of Characteristics distinguishing analysed species pairs ( species-groups according to canonical roots. baltica colorata polyphylloides L, LL, WL, LP,

¥ ¥ ¥ Least squared means with conﬁdence inter- . . .

Table 4. A. coccinea A. arenaria A. maritima A. macrocephala L, LL, WL, LP, A A A

vals of taxonomically important features were ANN. BOT. FENNICI Vol. 42 • Morphological variation in eight taxa of Anthyllis vulneraria 299 calculated by one-way ANOVA for reclassified of the species pairs were ratio of leaf length and species-groups. width, ratio of calyx length and width, and ratio of hypsophylls length and width. All other interval characteristics were suitable to distinguish at Results least one species pair. Most of the nominal and ordinal characteris- Results of the univariate GLM analysis of metric tics were statistically significant (Table 5). The and counted interval characteristics showed that Kruskal-Wallis test showed that only the shape four species, A. vulneraria, A. coccinea, A. ¥ bal- of rosette leaves failed to significantly distin- tica and A. ¥ polyphylloides, were distinguish- guish species. able from the other species (Table 4). However, Standard discriminant analysis of all char- some species remained indistinguishable with acteristics revealed that only 11 of them were respect to the diagnostic characteristics. Anthyl- statistically significant (Table 5), consisting of lis arenaria did not differ from A. maritima, A. six nonparametric, three counted interval, and macrocephala and A. ¥ colorata, A. maritima two metric interval characteristics. These signif- did not differ from A. macrocephala, and A. icant characteristics consist of apices of bracts; macrocephala did not differ from A. ¥ colorata. hairiness of petiole, calyx, and stem; colour of Characteristics that did not differ between any corolla and calyx; number of inflorescences,

Table 5. Results of the characteristics analysed with different methods. Critical p value is 0.05. Characteristic abbreviations as in Table 3.

Characteristics used p value of p value of p value of GLM p value of in analysis discriminant Kruskal-Wallis test for interval discriminant analysis test for characteristics analysis of nonparametric iteratively characteristics reclassiﬁed species

SR 0.0876 0.0010 0.1876 HLT < 0.0001 < 0.0001 < 0.0001 HSP < 0.0001 < 0.0001 < 0.0001 SRL 0.3341 < 0.1147 0.5813 HP < 0.0001 < 0.0001 < 0.0001 HST < 0.0001 < 0.0001 < 0.0001 CIF < 0.0001 < 0.0001 < 0.0001 DIF 0.4405 < 0.0001 0.0028 CP < 0.0001 < 0.0001 < 0.0001 HLB 0.1501 0.0178 0.2809 NS 0.0002 < 0.0001 < 0.0001 NIF < 0.0001 < 0.0001 < 0.0001 NSL 0.0632 < 0.0001 0.0056 NRL 0.0027 < 0.0001 0.1714 L 0.0090 < 0.0001 < 0.0001 LL 0.1967 < 0.0001 0.0469 WL 0.1784 < 0.0001 0.1382 LL/WL 0.1503 0.0603 0.1157 LP 0.5169 < 0.0001 0.0032 WP 0.2948 < 0.0001 0.0037 LP/WP 0.3238 0.5280 0.0018 LF 0.0002 0.0063 0.0046 LHL 0.7691 < 0.0001 0.0639 WHL 0.3083 < 0.0001 0.0135 LHL/WHL 0.2694 0.2307 0.1475 LS 0.2903 < 0.0001 0.7248 300 Puidet et al. • ANN. BOT. FENNICI Vol. 42

0 2

Root –2

–4 Anthyllis vulneraria A. coccinea A. arenaria –6 A. maritima A. macrocephala A. x baltica A. x colorata –8 A. x polyphylloides –10 –8 –6 –4 –2 0 2 4 6 Fig. 1. Scatterplot of canon- Root 1 ical discriminant analysis.

2 2 0 Root

–2

Anthyllis vulneraria –4 A. coccinea A. arenaria A. maritima –6 A. macrocephala Fig. 2. Scatterplot of A. x baltica canonical discriminant A. x colorata analysis for reclassified –8 A. x polyphylloides –10 –8 –6 –4 –2 0 2 4 6 8 species-clusters (by itera- Root 1 tive discriminant analysis). stems, and rosette leaves; height of plant; and pairs of species: A. vulneraria–A. ¥ baltica, A. length of corolla. Five metric interval charac- arenaria–A. maritima and A. macrocephala–A. ¥ teristics were statistically significant according polyphylloides. to GLM and discriminant analyses. These char- After iterative canonical analysis with deter- acteristics are number of stems, inflorescences mination correction, the ordination of canonical and rosette leaves, height of plant, and length of roots yielded a scatterplot, in which seven of the corolla. eight species formed distinctive clusters (Fig. 2). A scatterplot of canonical roots shows distin- Cluster edges overlap to some extent, but most guishable mono-specific groups of A. coccinea of the overlap is caused by the variability of A. and A. ¥ colorata (Fig. 1) and three overlapping maritima. ANN. BOT. FENNICI Vol. 42 • Morphological variation in eight taxa of Anthyllis vulneraria 301

A comparison of the initial and final classification cross-tabulation matrix revealed that A. 0 0 1.3 1.1 3.2 coccinea was classified 100% correctly (Table 0 0 31.8 6). Anthyllis arenaria, A. maritima, A. macro- polyphylloides

cephala and A. ¥ colorata were correctly clas- ¥ siﬁed > 90%, whereas 23.3% of A. vulneraria were classiﬁed as A. ¥ baltica and 3.4% as A. ¥ colorata. Anthyllis ¥ baltica was classifed 21.9% 3.4 0 0 0 0 1.0 0 97.4

A. vulneraria A. maritima colorata as and 6.7% as . The most poorly classified species was A. ¥ polyphyl- ¥ loides, of which only 31.8% were correctly classified. Half of these individuals were classified 0 0 2.2 0 as A. macrocephala, 9.1% as A. maritima and 0 0 23.3 69.4 baltica

9.1% as A. arenaria. ¥ Mean plant height, number of inﬂorescences,

number of stems, and leaf length differentiated species most successfully (Table 7). For exam- ple, A. coccinea is the shortest species (mean 0 0 0 2.2 0 2.6 93.6 height 10.7 ± 1.1 cm), whereas A. ¥ polyphyl- 50.0

loides is the tallest (mean height 41.2 ± 2.4 cm). macrocephala Mean heights of other species ranged between 17.2 cm and 27.9 cm. The variation patterns of the other characteristics were similar. Reclassiﬁed species 0 0 1.3 3.2 6.7 0 9.1 94.5

Discussion maritima

Several authors have suggested that Anthyllis 0 0 0 0 taxa are distinguishable by plant height, shape of 1.0 0 9.1 97.4

the hypsophyll apices, stem hairiness, and colour arenaria of corolla and calyx (Cullen 1968, Juzepczuk 1945, Eglite & Krall 1996, Krall 1999). These characteristics reliably distinguish the analysed 0 0 0 0 species according to our statistical analysis 0 0 0 (Table 5). The number of stems and inﬂores- 100 coccinea cences and hairiness of petiole and calyx, mentioned by Eglite and Krall (1996) and Krall

(1999), also statistically distinguish the analysed species. Easily measured characteristics, such as 0 0 0 0 0 0 73.3 number of rosette leaves and lengths of corolla 21.9 and calyx also reliably distinguish these species, vulneraria although they have not been mentioned in the literature. GLM analysis indicates that A. ¥ colorata does not differ from A. macrocephala. Anthyllis ¥ colorata is probably a hybrid between A. macrocephala and A. vulneraria (Juzepczuk 1945, Eglite & Krall 1996), but ﬁve characteristics clearly distinguish the second probable parent baltica colorata polyphylloides

species from A. ¥ colorata (Table 4). Anthyllis Table 6. Frequency matrix of analyzed species: the numbers are percentages species reclassiﬁ ed after analysis posterior probabilities. Initial classiﬁcation of species vulnereria coccinea arenaria maritima macrocephala ¥ ¥ ¥ 302 Puidet et al. • ANN. BOT. FENNICI Vol. 42

¥ colorata is also similar to A. maritima and A. arenaria, suggesting that these species might also be closely related. Interval characteristics 6.0 ± 0.9 4.8 ± 0.3 3.6 ± 1.8 4.3 ± 0.3 1.7 ± 0.1 2.6 ± 0.2 1.0 ± 0.0 0.4 ± 0.0 2.5 ± 0.2 0.4 ± 0.0 1.4 ± 0.1 0.3 ± 0.0 4.8 ± 0.7 5.0 ± 0.4 16.6 ± 1.6 41.2 ± 2.4 clearly distinguish the other four species. polyphylloides

. According to discriminant analysis, incor- A rectly classiﬁed individuals of A. vulneraria are

mostly classiﬁed as A. ¥ baltica (probably A. vulneraria ¥ A. maritima; Minjaev & Akulova 1987), and vice versa (Table 6). Its other parent colorata

4.0 ± 0.4 6.0 ± 0.8 2.9 ± 0.2 8.3 ± 0.9 3.1 ± 0.1 1.2 ± 0.1 2.6 ± 0.1 0.9 ± 0.0 0.3 ± 0.0 2.8 ± 0.1 0.4 ± 0.0 1.0 ± 0.0 0.2 ± 0.0 4.7 ± 0.3 3.6 ± 0.2

¥ ¥

. species, A. maritima, was not classiﬁed as A. baltica, but their similarity is revealed by seven individuals of A. ¥ baltica that were classi- ﬁed as A. maritima. According to this analysis, A. maritima, together with A. arenaria and baltica A ¥ 4.3 ± 0.3 6.2 ± 0.5 3.5 ± 0.1 7.6 ± 0.6 2.8 ± 0.1 1.0 ± 0.0 2.9 ± 0.1 0.8 ± 0.0 0.3 ± 0.0 2.8 ± 0.1 0.4 ± 0.0 1.0 ± 0.0 0.2 ± 0.0 5.6 ± 0.2 3.7 ± 0.1 A. macrocephala, are closely related to A. ¥ 19.6 ± 0.8 21.5 ± 1.2 A. polyphylloides. According to Krall (1999), A. ¥ polyphylloides is the hybrid of A. arenaria and A. macrocephala, but Minjaev and Akulova (1987) suggested that three species — A. arenaria ¥ A. macrocephala ¥ A. maritima — contribute to the 3.3 ± 0.4 5.9 ± 0.8 3.2 ± 0.2 5.1 ± 0.9 3.5 ± 0.1 1.2 ± 0.1 3.0 ± 0.1 0.9 ± 0.0 0.3 ± 0.0 2.7 ± 0.1 0.4 ± 0.0 1.1 ± 0.0 0.3 ± 0.0 4.3 ± 0.3 4.2 ± 0.2 origin of this species. In all analyses, only A. coccinea is distinctly A. macrocephala Species different from the others (Fig. 1), which sup-

ports the distinction of a species or subspecies, A. coccinea, from A. vulneraria s. lato. Strongly expressed morphological similarity between all other species indicates the delimitation of a A. maritima single species, A. vulneraria s. lato (Cullen 1968,

Hegi 1975, Lukaszewska et al. 1983c). The for- mation of subgroups (A. vulneraria–A. ¥ baltica, A. arenaria–A. maritima, A. macrocephala–A. ¥ colorata–A. ¥ polyphylloides) supports the idea A. arenaria of three subspecies (subsp. vulneraria, subsp. maritima and subsp. polyphylla) (Cullen 1968, Krall 1983). This study incorporated an original method 2.0 ± 0.4 4.1 ± 0.3 3.8 ± 0.3 2.3 ± 0.7 5.7 ± 0.6 5.5 ± 0.5 3.0 ± 0.2 3.5 ± 0.1 3.8 ± 0.1 5.2 ± 0.8 5.9 ± 0.6 6.5 ± 0.6 1.7 ± 0.1 3.3 ± 0.1 3.2 ± 0.1 0.6 ± 0.1 1.1 ± 0.0 1.1 ± 0.0 2.7 ± 0.1 3.0 ± 0.1 3.0 ± 0.1 0.8 ± 0.0 0.9 ± 0.0 0.9 ± 0.0 0.3 ± 0.0 0.3 ± 0.0 0.3 ± 0.0 2.7 ± 0.1 2.8 ± 0.1 2.8 ± 0.1 0.4 ± 0.0 0.4 ± 0.1 0.4 ± 0.0 0.8 ± 0.0 1.1 ± 0.0 1.0 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 4.9 ± 0.0 5.1 ± 0.2 5.3 ± 0.2 2.1 ± 0.2 for 4.2 ± 0.2 iterative 4.0 ± 0.1 improvement of the classiﬁcation

A. coccinea structure. The suitability of this approach is indicated by improved cluster interpretation and correction of initial misclassification. Canonical discriminant analysis reveals that most of the reclassified Anthyllis taxa can be differentiated 3.0 ± 0.3 4.4 ± 0.5 3.1 ± 0.1 6.0 ± 0.6 2.4 ± 0.1 0.8 ± 0.0 3.0 ± 0.1 0.8 ± 0.0 0.3 ± 0.0 2.9 ± 0.1 0.4 ± 0.0 0.9 ± 0.0 0.2 ± 0.0 5.2 ± 0.2 3.2 ± 0.1 17.2 ± 0.8 10.7 ± 1.1 23.1 ± 0.9 22.5 ± 0.8 reliably 27.9 ± 1.2 using morphological features (Fig. 2), A. vulneraria although all these species belong to the section Anthyllis (syn. Vulneraria DC.) (Juzepczuk 1945, Tabaka 1982), and are morphologically quite similar (Krall 1983). According to the statistical Mean values of interval characteristics for analyzed species (mean ± standard error), characteristic abbreviations as in Table 3. analysis, the similarity is expressed strongly for A. arenaria, A. maritima, and A. macrocephala Table 7. Characteristics NS NIF NSL NRL L LL WL LL/WL LP WP LP/WP LF LHL WHL LHL/WHL LS and for their hybrids, or reflects a specific trait ANN. BOT. FENNICI Vol. 42 • Morphological variation in eight taxa of Anthyllis vulneraria 303 of A. maritima, a highly variable species. It is Hultén, E. & Fries, M. 1986a: Atlas of North European vascular plants, north of the tropic of cancer, 2. — Koeltz highly questionable whether separation of these Sci. Books, Königstein. three species is justified, and this in turn does not Hultén, E. & Fries, M. 1986b: Atlas of North European vas- support differentiation of their hybrids. cular plants, north of the tropic of cancer, 3. — Koeltz Only molecular studies will clarify defini- Sci. Books, Königstein. tively the questions surrounding Anthyllis species Jalas, J. 1950: Zur Kausalanalyze der Verbereitung einiger nordischen Os- und Sandpflanzen. — Ann. Soc. Zool. taxonomy, and this work should be considered a Bot. Fennici 3: 123–127. framework for subsequent genetic analyses. Juzepczuk, S. 1945: Anthyllis L. — In: Komarov, V. (ed.), Flora USSR 11: 262–276. Leningrad & Moskva. Kalinowski, A., Szweykowski, J. & Kaczmarek, Z. 1983a: Acknowledgements Analysis of the variability of natural Anthyllis vulneraria s. lato populations from Baltic coast. Patterns of three enzyme systems following electrophoretic separation. We are grateful to Tatjana Oja for giving valuable assistance — Bull. Soc. des Amis des Sciences & des Lettres 23: during the preparation of the manuscript. We also thank 67–78. Ilmar Part and Robert Szava-Kovats for language correction Kalinowski, A., Szweykowski, J. & Kaczmarek, Z. 1983b: and we express our gratitude to the curators at the herbaria Analysis of the variability of natural Anthyllis vulner- for their help. We are very grateful to reviewers for valuable aria s. lato populations from Baltic coast. Patterns comments on the manuscript. 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